The known coronary artery disease risk factors can account for only half of the incidence of coronary artery disease; further, approximately 25 percent of these individuals have no identifiable risk factors. One might surmise, therefore, that there are one or more hitherto unrecognized risk factors. There is a growing feeling in the lipoprotein field that lipoprotein variants and/or ramnants may represent unrecognized risk factors. While evidence is accumulating to support this concept in animal models of atherosclerosis, the number of variants and their origin, composition, atherogenic potential and correlation with clinical history and known risk factors has not been established in humans. We have developed over the past 2-1/2 years a uniquely rapid, quantitative, high volume, high resolution method of lipoprotein isolation that makes possible the ability to test the variant lipoprotein hypothesis in a comprehensive manner. We are convinced, based upon extensive preliminary studies, that this system, single vertical spin (SVS) ultracentrifugation, is ready for final development and validation. We propose in this application to complete the validation of the SVS system and to subsequently develop a protocol for comprehensive multivariant lipoprotein analysis suitable for testing the hypothesis of altherogenic variant lipoproteins. We have evidence to suggest that the SVS system, in its various modifications, can isolate and quantitate VHDL, HDL3, HDL2a, HDL2b, MDL (Lp(a)), LDL, two populations of IDL and VLDL. A key feature of our proposed multivariant lipoprotein protocol is an automatic method of generation of cholesterol and triglyceride lipoprotein profiles (the vertical autoprofiler, VAP) that allows quantitation of these nine lipoprotein subfractions via computer-assisted curve deconvolution. The remainder of the multivariant protocol will involve the following analyses of the nine lipoprotein subfractions: comprehensive lipid analysis, radioimmunoassay for six apolipoproteins, gradient gel electrophoresis and electron microscopy (particle size) and strip electrophoresis (particle charge). Our goal is to achieve a highly reproducible and accurate system, supported by rigorous quality control, sufficient to allow the initiation of a pilot case-control search for potentially atherogenic variant lipoproteins in young, low risk males.